Thiophene-amd thiazolesulfonamides as antineoplastic agents

ABSTRACT

The present invention provides antineoplastic compounds of the formula (D).

[0001] In recent years fundamental advances have been made in thedevelopment of chemical agents and regimens of therapy to combatneoplastic diseases. Despite these continuing advances, cancers continueto exact intolerable levels of human pain and suffering. The need fornew and better methods of treating malignant neoplasms and leukemiascontinues to fuel efforts to create new classes of compounds, especiallyin the area of inoperable or metastatic solid tumors. The recentavalanche of information regarding the basic biological processesinvolved in neoplasms has led to a deeper understanding of theheterogeneity of tumors. It is because of this extreme heterogeneityamong populations of neoplastic cells that new chemotherapeutic agentsshould have a wide spectrum of activity and an acceptable therapeuticindex. In addition, such agents must be chemically stable and compatiblewith other agents. It is also important that any chemotherapeuticregimen be as convenient and painless as possible to the patient.

[0002] Chemotherapy and radiation are frequently used in the treatmentof cancer and, although they often produce some response in themalignant disease, they are rarely curative. Most solid tumors increasein mass through the proliferation of malignant cells and stromal cells,including endothelial cells. In order for a tumor to grow larger than2-3 millimeters in diameter, it must form a vasculature, a process knownas angiogenesis. Suppression of tumor-induced angiogenesis byangiostatin and endostatin has been reported to result in antitumoractivity (O'Reilly, et al., Cell, 88, 277-285 (1997)). Becauseangiogenesis is a critical component of the mass expansion of most solidtumors, the development of new agents for the inhibition of this processrepresents a promising approach for antitumor therapy. This approach toantitumor therapy may lack the toxic side effects or drugresistance-inducing properties of conventional chemotherapy (JudahFolkman, Endogenous inhibitors of Angiogenesis, The Harvey Lectures,Series 92, pages 65-82, Wiley-Liss Inc., (1998)).

[0003] The present invention provides novelN-[benzoyl]-heteroarylsulfonamide compounds useful in the treatment ofsusceptible neoplasms.

[0004] The present invention provides compounds of Formula I:

[0005] where:

[0006] —X═Y— is

[0007] R¹ is selected from the group consisting of halo, C₁-C₆ alkyl,and CF₃;

[0008] R² is selected from the group consisting of halo, —NO₂, C₁-C₆alkyl, and CF₃;

[0009] R³ is hydrogen, C₁-C₆ alkyl, C₁-C₄ alkoxy, C₁-C₆ alkylthio, orhalo;

[0010] R⁴ is selected from the group consisting of hydrogen, halo, C₁-C₄alkoxy, C₁-C₆ alkyl, —COO(C₁-C₆ alkyl), C₁-C₆ alkyl optionallysubstituted with C₁-C₄ alkoxy, cyano, C₁-C₆ alkylthio, CF₃, S-phenyl,and pyridinyl;

[0011] R⁵ is halo, C₁-C₆ alkyl, or C₁-C₄ alkoxy; or

[0012] a pharmaceutically acceptable base addition salt thereof.

[0013] The present invention further provides a method of treatingsusceptible neoplasms, in a mammal comprising administering to a mammalin need of such treatment an oncolytically effective amount of acompound of Formula I or a pharmaceutically acceptable base additionsalt thereof.

[0014] The present invention also provides a method of suppressing tumorangiogenesis in a mammal comprising administering to a mammal in need ofsuch treatment an angiogenesis suppressing amount of a compound ofFormula I or a pharmaceutically acceptable base addition salt thereof.

[0015] The present invention also provides a pharmaceutical formulationcomprising a compound of Formula I or a pharmaceutically acceptable baseaddition salt thereof and one or more pharmaceutically acceptableexcipients.

[0016] This invention also provides the use of a compound of Formula Ifor the manufacture of a medicament for the treatment of susceptibleneoplasms. Additionally, this invention provides a pharmaceuticalformulation for the treatment susceptible neoplasms containing acompound of Formula I with a pharmaceutically acceptable carrier orexcipient. Furthermore, this invention includes a method for thetreatment of susceptible neoplasms that comprises administering aneffective amount of a compound of Formula I.

[0017] The general chemical terms used in the formulae above have theirusual meanings. For example, the term “C₁-C₆ alkyl” includes methyl,ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl,pentyl, and hexyl moieties. The term “C₁-C₄ alkyl” is included withinthe meaning of C₁-C₆ alkyl and is taken to mean methyl, ethyl, propyl,isopropyl, butyl, sec-butyl, isobutyl and tert-butyl. The term “C₁-C₄alkoxy” is taken to mean a C₁-C₄ alkyl group linked to the parentmolecule through an oxygen atom, and includes the groups methoxy,ethoxy, isopropoxy, and the like. The term “halo” is taken to meanchlorine, bromine, fluorine, or iodine.

[0018] The term “mammal” is taken to mean any of various warm-bloodedvertebrate animals of the class Mammalia, most preferably humans,characterized by a covering of hair on the skin and, in the female,milk-producing mammary glands for nourishing the young.

[0019] While all of the compounds of Formula I are useful antineoplasticagents, certain classes of compounds are preferred. The followingparagraphs describe such preferred classes.

[0020] a) R¹ is halo, C₁-C₆ alkyl, or CF₃;

[0021] b) R¹ is chloro, bromo, fluoro, methyl, or CF₃;

[0022] c) R¹ is halo or C₁-C₆ alkyl;

[0023] d) R¹ is chloro;

[0024] e) R¹ is bromo;

[0025] f) R¹ is methyl;

[0026] g) R¹ is CF₃;

[0027] h) R² is halo, nitro, C₁-C₆ alkyl, or CF₃;

[0028] i) R² is chloro, bromo, nitro, methyl, or CF₃;

[0029] j) R² is halo or C₁-C₆ alkyl;

[0030] k) R² is chloro;

[0031] l) R² is bromo;

[0032] m) R¹ is methyl;

[0033] n) R² is NO₂;

[0034] o) R² is CF₃;

[0035] p) —X═Y— is

[0036] q) —X═Y— is

[0037] r) R³ is H, halo, C₁-C₆ alkyl, C₁-C₄ alkoxy, or C₁-C₆ alkylthio;

[0038] s) R³ is H, chloro, bromo, methyl, methoxy, or methylthio;

[0039] t) R³ is H or halo;

[0040] u) R³ is H;

[0041] v) R³ is bromo;

[0042] w) R³ is chloro;

[0043] x) R³ is C₁-C₆ alkyl;

[0044] y) R³ is methyl;

[0045] z) R³ is C₁-C₄ alkoxy;

[0046] aa) R³ is methoxy;

[0047] bb) R³ is C₁-C₆ alkylthio;

[0048] cc) R³ is methylthio;

[0049] dd) R⁴ is H, halo, C₁-C₆ alkyl, C₁-C₆ alkylthio, C₁-C₆ alkyloptionally substituted with C₁-C₄ alkoxy, C₁-C₄ alkoxy, cyano, S-phenyl,or pyridinyl;

[0050] ee) R⁴ is H, chloro, bromo, methyl, ethyl, propyl, methylthio,CH₂OCH₃, methoxy, cyano, S-phenyl, or pyridinyl;

[0051] ff). R⁴ is C₁-C₆ alkyl;

[0052] gg) R⁴ is methyl;

[0053] hh) R⁴ is ethyl;

[0054] ii) R⁴ is propyl;

[0055] jj) R⁴ is halo;

[0056] kk) R⁴ is chloro;

[0057] ll) R⁴ is bromo;

[0058] mm) R⁴ is hydrogen;

[0059] nn) R⁴ is C₁-C₄ alkoxy;

[0060] oo) R⁴ is methoxy;

[0061] pp) R⁴ is —COO(C₁-C₆ alkyl);

[0062] qq) R⁴ is C₁-C₆ alkyl optionally substituted with C₁-C₄ alkoxy;

[0063] rr) R⁴ is CH₂OCH₃;

[0064] ss) R⁴ is cyano;

[0065] tt) R⁴ is C₁-C₆ alkylthio;

[0066] uu) R⁴ is S-phenyl;

[0067] vv) R⁴ is pyridinyl;

[0068] ww) R⁵ is halo;

[0069] xx) R⁵ is chloro;

[0070] yy) R⁵ is C₁-C₄ alkoxy;

[0071] zz) R¹ is methoxy;

[0072] aaa) R⁵ is C₁-C₆ alkyl;

[0073] bbb) R⁵ is methyl;

[0074] ccc) R¹ and R² are independently halo or C₁-C₆ alkyl;

[0075] ddd) R¹ and R² are chloro, bromo, or R¹ is methyl and R² ischloro;

[0076] eee) R¹ and R² are chloro;

[0077] fff) R¹ is methyl and R² is chloro;

[0078] It will be understood that the above classes may be combined toform additional preferred classes.

[0079] The compounds of Formula I are antineoplastic agents. Thus, thepresent invention also provides a method of treating a susceptibleneoplasm in a mammal that comprises administering to a mammal in need ofsaid treatment an oncolytically effective amount of a compound ofFormula I. The present compounds are believed to be useful in treatingsusceptible neoplasms, including tumors and carcinomas, such asneoplasms of the central nervous system: glioblastoma multiforme,astrocytoma, oligodendroglial tumors, ependymal and choroid plexustumors, pineal tumors, neuronal tumors, medulloblastoma, schwannoma,meningioma, meningeal sarcoma; neoplasms of the eye: basal cellcarcinoma, squamous cell carcinoma, melanoma, rhabdomyosarcoma,retinoblastoma; neoplasms of the endocrine glands: pituitary neoplasms,neoplasms of the thyroid, neoplasms of the adrenal cortex, neoplasms ofthe neuroendocrine system, neoplasms of the gastroenteropancreaticendocrine system, neoplasms of the gonads; neoplasms of the head andneck: head and neck cancer, oral cavity, pharynx, larynx, odontogenictumors; neoplasms of the thorax: large cell lung carcinoma, small celllung carcinoma, non-small cell lung carcinoma, malignant mesothelioma,thymomas, primary germ cell tumors of the thorax; neoplasms of thealimentary canal: neoplasms of the esophagus, neoplasms of the stomach,neoplasms of the liver, neoplasms of the gallbladder, neoplasms of theexocrine pancreas, neoplasms of the small intestine, veriform appendixand peritoneum, adneocarcinoma of the colon and rectum, neoplasms of theanus; neoplasms of the genitourinary tract: renal cell carcinoma,neoplasms of the renal pelvis and ureter, neoplasms of the bladder,neoplasms of the urethra, neoplasms of the prostate, neoplasms of thepenis, neoplasms of the testis; neoplasms of the female reproductiveorgans: neoplasms of the vulva and vagina, neoplasms of the cervix,addenocarcinoma of the uterine corpus, ovarian cancer, gynecologicsarcomas; neoplasms of the breast; neoplasms of the skin: basal cellcarcinoma, squamous cell carcinoma, dermatofibrosarcoma, Merkel celltumor; malignant melanoma; neoplasms of the bone and soft tissue:osteogenic sarcoma, malignant fibrous histiocytoma, chondrosarcoma,Ewing's sarcoma, primitive neuroectodermal tumor, angiosarcoma;neoplasms of the hematopoietic system: myelodysplastic sydromes, acutemyeloid leukemia, chronic myeloid leukemia, acute lymphocytic leukemia,HTLV-1 and T-cell leukemia/lymphoma, chronic lymphocytic leukemia, hairycell leukemia, Hodgkin's disease, non-Hodgkin's lymphomas, mast cellleukemia; and neoplasms of children: acute lymphoblastic leukemia, acutemyelocytic leukemias, neuroblastoma, bone tumors, rhabdomyosarcoma,lymphomas, renal tumors. In particular, the present compounds arebelieved to be useful in treating solid tumors, especially tumors of thecolon and rectum. It is preferred that the mammal to be treated by theadministration of the compounds of Formula I is human.

[0080] The compounds of the present invention are acidic in nature andaccordingly may react with any of a number of inorganic and organicbases, for example, amines and quaternary ammonium bases, to formpharmaceutically acceptable base addition salts. It is preferable toconvert the compounds of Formula I to their pharmaceutically acceptablebase addition salts for ease of administration when aqueous solutions ofthe subject compound are required. The Formula I compounds can reactwith basic materials such as alkali metal- or alkaline earth metalhydroxides, carbonates, and bicarbonates including, without limitation,sodium hydroxide, sodium carbonate, potassium hydroxide, calciumhydroxide, and lithium hydroxide to form pharmaceutically acceptablesalts such as the corresponding sodium, potassium, lithium, or calciumsalt. The sodium and potassium salts are especially preferred.

[0081] Examples of amines suitable for forming salts are: primary,secondary and tertiary aliphatic and aromatic amines, such asmethylamine, ethylamine, propylamine, i-propylamine, the four isomericbutylamines, dimethylamine, diethylamine, diethanolamine, dipropylamine,diisopropylamine, di-n-butylamine, pyrrolidine, piperidine, morpholine,trimethylamine, triethylamine, tripropylamine, quinuclidine, pyridine,quinoline and isoquinoline, especially ethyl-, propyl-, diethyl- ortriethylamine, but particulary isopropylamine and diethanolamine.

[0082] Examples of quaternary ammonium bases are in general the cationsof hydroxyammonium salts, for example the tetramethylarumonium cation,the trimethylbenzylammonium cation, the triethylbenzylammonium cation,the tetraethylammonium cation or the trimethylethylammonium cation, butalso the ammonium cation.

[0083] The skilled artisan will appreciate that the introduction ofcertain substituents will create asymmetry in the compounds of FormulaI. The present invention contemplates all enantiomers and mixtures ofenantiomers, including racemates. It is preferred that the compounds ofthe invention containing chiral centers are single enantiomers. Thepresent invention further contemplates all diastereomers.

[0084] The compounds of the present invention can be prepared by avariety of procedures, some of which are illustrated in the schemesbelow. It will be recognized by one of skill in the art that theindividual steps in the following schemes may be varied to provide thecompounds of Formula I. Some of these variations are discussed.

[0085] The particular order of steps required to produce the compoundsof Formula I is dependent upon the particular compound beingsynthesized, the starting compound, and the relative lability of thesubstituted moieties.

[0086] The compounds of the present invention may be prepared by methodswell known to one of ordinary skill in the art. Generally, the compoundsof Formula I can be prepared by coupling an appropriately substitutedthienyl- or thiazolyl-sulfonamide with an appropriately substitutedbenzoic acid as illustrated in the following schemes. The variables R¹,R², X, and Y are as previously defined.

[0087] The optionally substituted benzoic acid is coupled to anappropriate sulfonamide under standard peptide coupling conditions wellknown to the skilled artisan. Specifically, the thienyl-or thethiazolyl-sulfonamides and the benzoic acid are coupled in the presenceof a peptide coupling reagent, optionally in the presence of a catalyst.Suitable peptide coupling reagents include N,N′-carbonyldiimidazole(CDI), N,N′-dicyclohexylcarbodiimide (DCC),1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC), and1-(3-(1-pyrrolidinyl)propyl)-3-ethylcarbodiimide (PEPC). Polymersupported forms of EDC (Tetrahedron Letters, 34(48), 7685 (1993)) andPEPC (U.S. Pat. No. 5,792,763) have been described, and are very usefulfor the preparation of the compounds of the present invention. Suitablecatalysts for the coupling reaction includeN,N-[dimethyl]-4-aminopyridine (DMAP). All of the reagents are combinedin a suitable solvent, typically dichloromethane, chloroform,tetrahydrofuran, dioxane, or diethyl ether and are stirred for from 1 to72 hours at a temperature of from ambient to about the refluxtemperature of the solvent. Where excess or unreacted sulfonamide orbenzoic acid remains in the reaction mixture, it may be removed by theaddition of an appropriate acidic or basic resin, followed byfiltration. Alternatively, these reagents may be removed by extractivetechniques. The desired product may be isolated by standard extractiveand crystallization techniques, and purified by chromatography orcrystallization as necessary or desired. Where polymer-bound reagentsare employed, they may be conveniently removed from the reaction mixtureby filtration.

[0088] The requisite benzoic acids and sulfonamides are eithercommercially available or may be prepared by methods well known to theskilled artisan, such as in the following synthetic schemes. Thevariables R¹, R², X, and Y are as previously defined and Z is a cyanogroup or a halide.

[0089] Synthetic Scheme II depicts sulfonylation of the thiophenes andthiazoles of formula (3) in the formation of the sulfonamides of formula(1). Synthetic conditions for sulfonylations are dependent on thefunctional groups of the thiophene starting material. For example, in(step a), a lithium base such as n-butyl lithium is used to create theanion of formula (4) in situ, at a temperature range of −78° C. to roomtemperature. The anion is quenched with a sulfonating reagent, such assulfur dioxide, (step b) to give compounds of formula (5). Formula (5)can be further reacted with N-chlorosuccinimide, (step c), to affordsulfonyl chlorides of formula (6). Alternatively, formula (4) may betreated with sulfuryl chloride, (step e) to give the sulfonyl chloridesof formula (6) directly (Howbert, J. J.; Mohamadi, F.; Spees, M. M.European Patent 0 467 613 A1). The skilled artisan will also appreciatethat the sulfonyl chloride of formula (6) may be prepared by thereaction of formula (3) with chlorosulfonic acid (step g). The sulfonylchlorides of formula (6) can be contacted with ammonium hydroxide, (stepd), to give the sulfonamides of formula (1)(Cremlyn, R. J.; Bassin, J.P. Farouk, S.; Potterton, M.; Mattu, T. Phosphorus, Sulfur SiliconRelat. Elem. 1992, 73 (1-4), 107-120); Besterman, J. M.; Delorme, D.;Rahil, J. WO 01 02411, 2001). Alternatively, formula (5) can be treatedwith hydroxylamine-O-sulfonic acid, (step f), to give sulfonamides offormula (1) directly (Mohamadi, F.; Spees, M. M. U.S. Pat. No.5,169,860).

[0090] The synthetic conditions of Synthetic Scheme II are well knownand appreciated in the art (J. Med. Chem., Graham, S. L., et al., 1989,32, 2548-2554; J. Med. Chem., Barnish, I. T. et al., 1981, 24, 959; J.Chem. Soc., Cymerman-Craig, J., et al., 1956, 4115).

[0091] The preparation of the requisite benzoic acids (2) maybeaccomplished by functional transformations well known to the skilledartisan as illustrated in Synthetic Scheme III. For example, when Z is acyano group the conversion to the carboxylic acid can be achieved underacidic conditions (Larock, R. C., Comprehensive Organic Transformations,2^(nd) Ed., copyright 1999, John Wiley & Sons, pp 1986-1987). When Z isa halide a metal promoted carbonylation can be performed with palladiumacetate and carbon monoxide in methanol to give the methyl benzoate (Id.at 1685-1687), then followed by a hydrolysis to afford the benzoic acidsof formula (2) (Id. at 1959-1968). One skilled in the art willappreciate further manipulation of the R groups of the startingcompounds of formula (3) and (8) that are done by known syntheticinterconversions such as an amino derivative to the corresponding halide(Id. at 677-679), a halide exchange with a metal-alkoxide (Id. at893-894) or a nucleophilic addition of appropriate sulfur or nitrogennucleophiles (Id. at 779-780).

[0092] The skilled artisan will also appreciate that not all of thesubstituents in the compounds of Formula I will tolerate certainreaction conditions employed to synthesize the compounds. These moietiesmay be introduced at a convenient point in the synthesis, or may beprotected and then deprotected as necessary or desired. Furthermore, theskilled artisan will appreciate that in many circumstances, the order inwhich moieties are introduced is not critical.

[0093] The following preparations and examples further illustrate thepreparation of compounds of the present invention and should not beinterpreted in any way as to limit the scope. Those skilled in the artwill recognize that various modifications may be made while notdeparting from the spirit and scope of the invention. All publicationsmentioned in the specification are indicative of the level of thoseskilled in the art to which this invention pertains.

[0094] The terms and abbreviations used in the instant Preparations andExamples have their normal meanings unless otherwise designated. Forexample “° C.”, “N”, “mmol”, “g”, “mL” “M”, “HPLC”, “IR”, “MS(FD)”,“MS(IS)”, “MS(FIA)”, “MS(FAB)”, “MS(EI)”, “MS(ES)”, “UV”, “TLC” and “¹HNMR”, refer to degrees Celsius, normal or normality, millimole ormillimoles, gram or grams, milliliter or milliliters, molar or molarity,high performance liquid chromatography, infra red spectrometry, fielddesorption mass spectrometry, ion spray mass spectrometry, flowinjection analysis mass spectrometry, fast atom bombardment massspectrometry, electron impact mass spectrometry, electron spray massspectrometry, ultraviolet spectrometry, thin layer chromatography andproton nuclear magnetic resonance spectrometry respectively. Inaddition, the absorption maxima listed for the IR spectra are only thoseof interest and not all of the maxima observed.

PREPARATION 1 5-(Methylthio)thiophene-2-sulfonamide

[0095]

[0096] 1.3 M n-Butyllithium in tetrahydrofuran (10 mL, 12.5 mmol;Aldrich) is added to a cold solution (−78° C.) of the2-(methylthio)thiophene (10.0 mmol; Aldrich) in anhydroustetrahydrofuran (5.0 mL/mmol). The mixture is allowed to react for 90min under nitrogen atmosphere. Sulfur dioxide is bubbled through thesolution for 30 min at −78° C. The mixture is warmed to room temperatureand concentrated by rotary evaporation. The residue is treated with asolution of sodium acetate (8 eq.) and hydroxylamine-O-sulfonic acid(2.5 eq) in water (4 mL/mol) and stirred at 25° C. for 1 hr. Thereaction mixture is made basic by addition of 1.0 N sodium hydroxide topH 10 and is extracted with diethyl ether (2×50 mL). The aqueous phaseis acidified to pH 2 with conc. hydrochloric acid and extracted withmethylene chloride (2×50 mL). The combined organic phases are washedwith saturated sodium bicarbonate (3×25 mL) and brine (50 mL), dried(sodium sulfate), filtered, and concentrated by rotary evaporation. Thecrude solid is purified by column chromatography with a mixturehexane/ethyl acetate (2:1) as the eluent. ¹H NMR (300 MHz), CDCl₃) δ:7.52 (d, 1H), 6.94 (d, 1H), 5.10 (br s, 2H), 2.58 (s, 3H).

PREPARATION 2 5-(Ethyl)thiophene-2-sulfonamide

[0097]

[0098] A solution of 2-ethylthiophene (1.78 mmol) dissolved inchloroform (1 mL/mmol) is added to a cold solution (0° C.) ofchlorosulfonic acid (0.35 mL, 5.35 mmol) in chloroform (1.3 mL/mmol).The mixture is stirred for 3 hr at room temperature with a drying tubeconnected.

[0099] The mixture is then poured over a cold mixture ofchloroform/water and stirred 10 min. The organic layer is washed withwater, dried over sodium sulfate and concentrated in vacuo. Two mL of anaqueous solution of ammonium hydroxide is added to the crude oil and themixture is stirred for 30 min. Solvent is concentrated in vacuo. Theresidue is employed without further purification. ¹H NMR (200 MHz,CDCl₃) δ: 7.48 (d, 1H, J=3.6 Hz), 6.74 (dd, 1H, J=3.7 Hz, 0.8 Hz), 5.2(br s, 2H), 2.9 (q, 2H, J=7.5 Hz), 1.32 (t, 3H, J=7.5 Hz).

PREPARATION 3 5-(Propyl)thiophene-2-sulfonamide

[0100]

[0101] A method similar to PREPARATION 2, with an exception for2-n-propylthiophene, is used. ¹H NMR (200 MHz, CDCl₃) δ: 7.46 (d, 1H,J=3.8 Hz), 6.72 (dd, 1H, J=3.8 Hz, 0.8 Hz), 5.30 (bs, 2H), 2.79 (t, 2H,J=7.4 Hz), 1.69 (q, 2H, J=7.4 Hz), 0.97 (t, 3H, J=7.4 Hz).

PREPARATION 4 5-(Methoxy)thiophene-2-sulfonamide

[0102]

[0103] 1.6 M n-butyllithium (1 mL, 1.75 mmol) is added to a coldsolution (−78° C.) of 2-methoxythiophene (1.75 mmol) in anhydroustetrahydrofuran (2.6 mL/mmol). The mixture is allowed to react for 45min under a nitrogen atmosphere. The solution is then warmed to 0° C.and sulfur dioxide is bubbled through the solution for 15 min and thenthe mixture is purged with nitrogen. The solvent is removed in vacuo andthe crude oil is dissolved in anhydrous methylene chloride (1 mL/mmol)and N-chlorosuccinimide is added (1.75 mmol). The mixture is stirred for2 hr at room temperature under a nitrogen atmosphere. It is filtered andthen concentrated in vacuo. The crude oil is dissolved in acetone (3mL/mmol) and 2 mL of an aqueous solution of ammonium hydroxide is added.The solution is stirred overnight. The solvent is concentrated in vacuo.The residue is dissolved in ethyl acetate and washed with water andbrine, dried over sodium sulfate, and concentrated under vacuum. Theresidue is purified by column chromatography with a mixture hexane/ethylacetate (7:3) as the eluent. ¹H NMR (200 MHz, CDCl₃) δ: 7.37 (d, 1H,J=4.3 Hz), 6.17 (d, 1H, J=4.3 Hz), 4.9 (br s, 2H), 3.94 (s, 3H).

PREPARATION 5 5-(Methyl)thiophene-2-sulfonamide

[0104]

[0105] A method similar to PREPARATION 2, with an exception for2-(methyl)thiophene, is used. ¹H NMR (200 MHz, CDCl₃) δ: 7.44 (d, 1H,J=3.7 Hz), 6.71 (br d, 1H, J=3.7 Hz), 4.92 (br s, 2H), 2.51 (d, 3H,J=0.9 Hz).

PREPARATION 6 5-(Methoxymethyl)thiophene-2-sulfonamide

[0106]

[0107] 2-(Hydroxymethyl)thiophene (4.4 mmol; Aldrich), silver (I) oxide(6.6 mmol, 1.5 eq; Aldrich) and methyl iodide (2.2 mmol, 5 eq; Aldrich)are dissolved in methylene chloride (2 mL/mmol) and stirred at roomtemperature for 48 hr. The mixture is filtered through celite and thesolvent is evaporated in vacuo. The residue is purified by columnchromatography with a mixture hexane/ethyl acetate (75:25) as theeluent.

[0108] 1.6 M N-butyllithium in tetrahydrofuran (0.6 mL, 0.9 mmol;Aldrich) is added to a cold solution (−78° C.) of the above product,2-(methoxymethyl)thiophene (0.87 mmol) in anhydrous tetrahydrofuran (1.3mL/mmol). The mixture is allowed to react for 30 min under a nitrogenatmosphere and is transferred via canula over a solution of sulfurylchloride (0.1 mL, 1.7 mmol; Aldrich) in hexane (2.5 mL/mmol). Thesolution is stirred under a nitrogen atmosphere for 2 hr and warmed toroom temperature. The mixture is diluted with ethyl acetate and washedwith water and brine, dried over sodium sulfate, and concentrated invacuo. The residue is dissolved in acetone (3 mL/mmol) and 2 mL of anaqueous solution of ammonium hydroxide is added with the solutionstirred overnight. The solvent is concentrated in vacuo. The residue isdissolved in ethyl acetate and washed with water and brine, dried oversodium sulfate, and concentrated under vacuum. The residue is purifiedby column chromatography with a mixture hexane/ethyl acetate (7:3) asthe eluent. ¹H NMR (200 MHz, CDCl₃) δ: 7.52 (d, 1H, J=3.7 Hz), 6.92 (d,1H, J=3.7 Hz), 5.23 (br s, 2H), 4.60 (s, 2H), 3.41 (s, 3H).

PREPARATION 7 4,5-Dibromothiophene-2-sulfonamide

[0109]

[0110] Phosphorus pentachloride (0.16 g, 0.8 mmol) is added portionwisewith stirring to chlorosulfonic acid (0.14 g, 1.2 mmol) and theresultant solution is cooled to 0° C., under a nitrogen atmosphere.2,3-Dibromothiophene (0.24 g, 0.8 mmol) is added with stirring and theresultant mixture is heated to 50° C. for 1 hr. Ice-water is added tothe reaction mixture and then it is extracted with ethyl acetate (20mL). The organic layer is concentrated and re-dissolved in acetone (5mL). Ammonium hydroxide (5 mL, concentrated) is added and the resultingmixture stirred for 30 min at room temperature. Brine (10 mL) and ethylacetate (20 mL) are added, the organic layer is separated, and theaqueous layer is extracted one more time with ethyl acetate (10 mL). Thecombined organic layers are dried over sodium sulfate, concentrated invacuo, and then chromatographed on silica (0.5% methyl alcohol inmethylene chloride) to give the title compound (58% yield) as a brownsolid. ES(−)MS m/z 318, (M−H)⁻ consistent with 2 Br.

PREPARATION 8 5-(Cyano)thiophene-2-sulfonamide

[0111]

[0112] A mixture of 5-bromothiophene-2-sulfonamide (0.50 g, 2.1 mmol),zinc cyanide (0.25 g, 2.1 mmol),tetrakis(triphenylphosphine)palladium(0) (0.072 g, 0.06 mmol) indimethylformamide (5 mL, anhydrous) is placed under microwave radiation(under nitrogen atmosphere, 160° C.) for 15 min. Thin layerchromatography (5% methyl alcohol in methylene chloride) demonstratesthe reaction is incomplete. Additionaltetrakis-(triphenylphosphine)palladium(0) (0.24 g, 0.2 mmol) anddimethylformamide (10 mL) are added to the reaction mixture and placedunder microwave radiation (under nitrogen atmosphere at 160° C.) for 37min. 10 mL of water and 20 mL of ethyl acetate are added to the reactionmixture. The organic phase is separated and the aqueous layer isextracted with 20 mL ethyl acetate. The combined organic layers aredried over sodium sulfate, concentrated in vacuo, and thenchromatographed on silica (0-5% methyl alcohol in methylene chloride) togive the title compound, as a white solid (0.22 g, 57% yield). ES(−)MSm/z 187, (M−H)⁻.

PREPARATION 9 5-(Methoxycarbonyl)thiophene-2-sulfonamide

[0113]

[0114] A mixture of 5-bromothiophene-2-sulfonamide (0.50 g, 2.1 mmol),triethyl amine (1 mL), methanol (1 mL), palladium acetate (0.046 g, 2.1mmol) and 1,3-bis(diphenylphosphino) propane (0.085 g, 2.1 mmol)(addition in that order) in dimethylformamide (5 mL, anhydrous) issaturated with carbon monoxide gas, at room temperature. This reactionmixture is heated to 100° C. and stirred overnight, under a carbonmonoxide atmosphere. 10 mL of brine and 10 mL of ethyl acetate are addedto the reaction mixture. The organic phase is separated and the aqueouslayer is extracted with 10 mL ethyl acetate. The combined organic layersare dried over sodium sulfate, concentrated in vacuo, and thenchromatographed on silica (0-1% methyl alcohol in methylene chloride) togive the title compound, as a yellow solid (0.15 g, 34% yield). ES(−)MSm/z 220, (M−H)⁻.

PREPARATION 10 2-Chlorothiazole-5-sulfonamide

[0115]

[0116] A method similar to PREPARATION 4, with an exception for2-chlorothiazole, is used.

PREPARATION 11 2-Methoxythiazole-5-sulfonamide

[0117]

[0118] A method similar to PREPARATION 1, with an exception for2-methoxy-thiazole, is used.

PREPARATION 12 2-Isopropyl-2,5-dihydrothiazole

[0119]

[0120] A solution of 1,4-dithiane-2,5-diol (20 g, 131 mmol) is suspendedin Et₂O (80 mL) in a round bottom flask that is equipped with acondenser and a gas inlet tube, Isobutyraldehyde (40 mL) and Na₂SO₄ (12g) are added then ammonia is bubbled through the reaction mixture for 20min at room temperature and 10 min at reflux. The reaction is thencooled to room temperature and the Na₂SO₄ is filtered and the solvent isdistilled at atmospheric pressure. The residue is distilled via avigreaux column at 130° C. at 7 in/Hg to afford the title compound (13.4g, 40%).

[0121] ES(+)MS mz/z 130, (M+H)⁺.

PREPARATION 13 2-Isopropylthiazole

[0122]

[0123] A solution of 2-isopropyl-2,5-dihydrothiazole (12.4 g, 95.9 mmol)in benzene (125 mL) is added to a solution of p-chloranil (23.6 g, 95.6mmol). The reaction mixture is refluxed for 2 hr and cooled to roomtemperature. A solution of 2 M NaOH (200 mL) is added and the reactionis stirred for 5 min then poured into a separators funnel. The organiclayer is separated and washed with 2 M NaOH (200 mL) and H₂O (2×100 mL).The aqueous layers are re-extracted with benzene and the organic layersare combined. Benzene is distilled off at atmospheric pressure to leavean oily residue which is distilled via a vigreaux column at 110° C. at 8in/Hg to provide the title compound (6.13 g, 48%) as a colorless oil.

[0124] ES(+)MS m/z 128, (M+H)⁺.

PREPARATION 14 2-Isopropylthiazole-5-sulfonamide

[0125]

[0126] To a solution of 2-isopropylthiazole (2 g, 15.7 mmol) in Et₂O (75mL) at −78° C. is added n-BuLi (12.8 mL of 1.6 M in hexanes, 20.4 mmol)dropwise (a pink precipitate is observed). After 40 min, the reactionmixture is warmed to 0° C. for 10 min then re-cooled to −78° C. Sulfurdioxide is bubbled over the surface of the reaction mixture for 5 min.The reaction mixture is warmed to room temperature and stirred for anadditional 2.5 hr. The reaction is cooled to 0° C. andN-chlorosuccinimide (4.20 g, 32.4 mmol) is added and the reaction isstirred for 1.5 hr. The reaction mixture is then filtered and theprecipitate is washed with Et₂O. The filtrate is concentrated undervacuum to give crude sulfonyl chloride which is dissolved in acetone (20mL) and added to a stirred solution of concentrated NH₄OH (20 mL) inacetone (50 mL) at 0° C. The reaction mixture is stirred for 5 min andthen partitioned between EtOAc and H₂O. The aqueous layer is separatedand extracted with EtOAc (2×). The organic layers are combined, dried(MgSO₄), filtered and evaporated under reduced pressure. The crudeproduct is recrystallized from CH₂Cl₂/acetone/hexanes to afford thetitle compound (1.89 g, 58%).

[0127] ES(+)MS m/z 207, (M+H)⁺.

PREPARATION 15 2-Methylthiazole

[0128]

[0129] To a stirred solution of 2-bromothiazole (5.0 g, 30.5 mmol) inEt₂O (60 mL) at −78° C. under nitrogen is added dropwise n-BuLi (14.6 mLof 1.6 M in hexanes, 36.6 mmol). The reaction mixture is stirred for 40min then dimethyl sulfate (4.75 mL, 50.3 mmol) is added dropwise and thereaction mixture is warmed to −10° C. (placed in a refrigerator) andleft standing overnight. The reaction is warmed to 0° C. and cautiouslyquenched with 2 M HCl (40 mL). The organic layer is separated andextracted with 2 M HCl (2×). The acid extracts are combined and madestrongly alkaline with 2 M NaOH and extracted with Et₂O (4×). Thecombined organic extracts are dried over KOH and the solvent isdistilled off at atmospheric pressure then the title compound isdistilled off at 128-130° C. (1.5 g, 49%).

[0130] ES(+)MS m/z 100, (M+H)⁺.

PREPARATION 16 2-Methylthiazole-5-sulfonamide

[0131]

[0132] To a stirred solution of n-BuLi (12.1 mL of 1.6 M in hexanes,19.4 mmol) in Et₂O (70 mL) at −78° C. under nitrogen is added dropwise asolution of 2-methyl-thiazole (1.48 g, 14.9 mmol) in Et₂O (70 mL). Thereaction mixture is stirred at −78° C. for 40 min then warmed to −20° C.Sulfur dioxide is bubbled over the solution for 5 min then the reactionis allowed to warm to room temperature overnight. N-Chlorosuccinimide(3.99 g, 29.9 mmol) is added and the reaction mixture allowed to stirfor 1 hr. The reaction is filtered and the filtrate concentrated undervacuum to provide the crude product. The crude product is dissolved inacetone (30 mL) and concentrated NH₄OH (20 mL) is added and the mixturestirred for 15 min. The reaction mixture is partitioned between EtOAcand H₂O. The aqueous layer is extracted with EtOAc (2×) and the organiclayers are combined, dried (MgSO₄), filtered and concentrated undervacuum.

[0133] Flash chromatography on silica gel eluting with a gradient [Hexto Hex:EtOAc (1:1)] provides the title compound (282 mg, 11%) as a tansolid.

[0134] ES(−)MS m/z 177, [M−H]⁻.

PREPARATION 17 2-Bromo-3-chlorothiophene

[0135]

[0136] To a solution 3-chlorothiophene (5.0 g, 42 mmol) in a mixture ofCHCl₃ (50 mL) and AcOH (50 mL) is added N-bromosuccinimide (8.3 g, 46mmol). The solution is heated to 50° C. After 1.5 hr, the reactionmixture is cooled to room temperature. Brine (100 mL) and Et₂O (200 mL)are added to the reaction mixture and the aqueous layer is extractedwith Et₂O (100 mL). The combined organic extracts are washed withsaturated NaHCO₃ then dried (Na₂SO₄), filtered and concentrated in vacuoto afford the title compound (5.4 g, 65%).

[0137]¹H NMR (300 MHz, CD₃OD) δ 6.94 (d, J=5.8 Hz, 1H), 7.50 (d, J=5.8Hz, 1H)

PREPARATION 18 5-Bromo-4-chlorothiophene-2-sulfonamide

[0138]

[0139] To phosphorous pentachloride (4.6 g, 22.2 mmol) is addedchlorosulfonic acid (2.2 mL, 33.3 mmol) under a nitrogen atmosphere. Thesolution is cooled to 0° C. and 2-bromo-3-chlorothiophene (1.0 g, 5.0mmol) is added. The mixture is heated to 50° C. for 1 hr. The reactionis cooled then quenched with ice/water and the solution is extractedwith CH₂Cl₂ (200 mL), and then the CH₂Cl₂ is removed under reducedpressure. The residue is dissolved in acetone (30 mL) and added to asolution of 29% NH₄OH (40 mL) in acetone (100 mL) at 0° C. The reactionmixture is stirred for 0.5 hr then the acetone is removed under reducedpressure. The residue is extracted with EtOAc (200 mL). The organiclayer is washed with brine then dried (Na₂SO₄), filtered andconcentrated in vacuo to afford the title compound (8.1 g, >100%), whichis used without further purification.

[0140] ES(−)MS m/z 274, [M−H]⁻ consistent with 1 Br and 1 Cl.

PREPARATION 19 4-Chlorothiophene-2-sulfonamide

[0141]

[0142] To a stirred solution of 5-bromo-4-chlorothiophene-2-sulfonamide(2.4 g,

[0143] 8.7 mmol) in AcOH (20 mL) is added zinc dust (1.7 g, 26.0 mmol).The reaction mixture is heated to 120° C. for 6 hr. After 6 hr, themixture is filtered and neutralized with 1 M NaOH. The aqueous layer isextracted with EtOAc (2×100 mL). The combined organic extracts are dried(Na₂SO₄), filtered and concentrated in vacuo. The crude product ischromatographed on silica gel, eluting with CH₂Cl₂ to afford the titlecompound (0.88 g, 52%).

[0144]¹H NMR (300 MHz, CD₃OD) δ 7.48 (s, 1H), 7.58 (s, 1H)

PREPARATION 20 2-Bromo-3-methylthiophene

[0145]

[0146] 3-Methylthiophene (5.0 g, 50.9 mmol) is dissolved in a solutionof CHCl₃ (50 mL) and AcOH (50 mL). N-Bromosuccinimide (9.5 g, 53.5 mmol)is added to the solution and the mixture is heated to 50° C. After 1.5hr, the reaction mixture is cooled to room temperature. Brine (100 mL)and Et₂O (200 mL) are added to the reaction mixture. The organic layeris separated and washed with 1 M NaOH and brine then dried (Na₂SO₄),filtered and concentrated in vacuo to give the title compound (6.4 g,71%) as a clear oil.

[0147]¹H NMR 300 MHz (CD₃OD) δ 2.14 (s, 3H), 6.81 (d, J=5.6 Hz, 1H),7.28 (d, J=5.6 Hz, 1H)

PREPARATION 21 5-Bromo-4-methylthiophene-2-sulfonamide

[0148]

[0149] To phosphorous pentachloride (6.5 g, 31 mmol) is addedchlorosulfonic acid (3.1 mL, 46.4 mmol). The mixture is cooled to 0° C.and 2-bromo-3-methylthiophene (5.4 g, 31 mmol) is added. The reactionmixture is heated to 50° C. for 1 hr. The reaction is cooled/quenchedwith ice/water and the solution extracted with CH₂Cl₂ (200 mL). Theorganic layer is washed with brine, dried (Na₂SO₄), filtered andconcentrated in vacuo. The residue is dissolved in acetone (20 mL) andadded to a solution of 29% NH₄OH (54 mL) in acetone (250 mL). Thereaction mixture is stirred for 0.5 hr then the acetone is removed underreduced pressure. The residue is extracted with EtOAc (2×100 mL). Thecombined organic extracts are washed with brine, dried (Na₂SO₄),filtered and concentrated in vacuo. The crude product is chromatographedon silica gel, eluting with CH₂Cl₂ to afford the title compound (5.3 g,58%).

[0150]¹H NMR (300 MHz, CD₃OD) δ 2.20 (s, 3H), 7.32 (s, 1H)

PREPARATION 22 4-Methylthiophene-2-sulfonamide

[0151]

[0152] To a stirred solution of 5-bromo-4-methylthiophene-2-sulfonamide(3.1 g, 12.1 mmol) in AcOH (30 mL) is added zinc dust (2.4 g, 36.2mmol). The reaction mixture is heated to reflux for 8 hr. After 8 hr,the reaction mixture is cooled and filtered. The filtrate is neutralizedwith 1 M NaOH. The aqueous layer is extracted with EtOAc (300 mL). Theorganics are dried (Na₂SO₄), filtered and concentrated in vacuo. Thecrude product is chromatographed on silica gel, eluting with CH₂Cl₂ toafford the title compound (0.90 g, 43%).

[0153]¹H NMR (300 MHz, CD₃OD) δ 2.26 (s, 3H), 7.27 (s, 1H), 7.41 (s, 1H)

PREPARATION 23 2-Trimethylsilyl-3-methoxythiophene

[0154]

[0155] A solution of n-BuLi (19.7 mL of 1.6 M in hexanes, 31.5 mmol) isadded dropwise to a solution of 3-methoxythiophene (3.0 g, 26.3 mmol) inanhydrous Et₂O (20 mL) under nitrogen at −70° C. The mixture is stirredat −70° C. for 2 hr. Chlorotrimethylsilane (4.5 mL, 35.4 mmol) is addedslowly to the solution. The mixture is warmed to room temperature andstirred for 3 hr. The reaction is quenched with water (50 mL) andhexanes (100 mL). The aqueous layer is extracted with hexanes (50 mL).The combined organic extracts are dried (Na₂SO₄), filtered andconcentrated. The crude product is chromatographed on silica gel,eluting with hexanes to afford the title compound (4.0 g, 82%) as acolorless liquid.

[0156]¹H NMR 300 MHz (CD₃OD) δ 0.29 (s, 9H), 3.81 (s, 3H), 6.92 (d,J=4.9 Hz, 1H), 7.40 (d, J=4.9 Hz, 1H)

PREPARATION 24 5-Trimethylsilyl-4-methoxythiophene-2-sulfonamide

[0157]

[0158] A solution of n-BuLi (11.8 mL of 2.5 M in hexanes, 29.4 mmol) isadded dropwise to a solution of 2-trimethylsilyl-3-methoxythiophene(2.19 g, 11.8 mmol) in anhydrous THF (40 mL) under nitrogen at −70° C.The mixture is stirred at −70° C. for 4 hr then sulfur dioxide isbubbled through the solution for 5 minutes. After stirring 2.5 hr,N-chlorosuccinimide (3.15 g, 23.6 mmol) is added to the suspension. Themixture is warmed to room temperature and stirred for 1 hr then thereaction mixture is filtered and the solids are washed with CH₂Cl₂. Thefiltrate is concentrated and the residue is dissolved in CH₂Cl₂ (200mL). The organic layer is washed with brine then dried (Na₂SO₄),filtered and concentrated. The residue is dissolved in acetone (20 mL)and added to a solution of 29% NH₄OH (20 mL) in acetone (30 mL) at 0° C.The mixture is stirred at 0° C. for 30 min then the acetone is removedunder reduced pressure and the residue is extracted with EtOAc (2×100mL). The organic extracts are washed with brine then dried (Na₂SO₄),filtered and concentrated. The crude product is chromatographed onsilica gel, eluting with Hex:EtOAc (3:1) to afford the title compound(0.77 g, 25%).

[0159]¹H NMR 300 MHz (CD₃OD) δ 0.29 (s, 9H), 3.31 (s, 3H), 7.49 (s, 1H)

PREPARATION 25 4-Methoxythiophene-2-sulfonamide

[0160]

[0161] To a solution of5-trimethylsilyl-4-methoxythiophene-2-sulfonamide (770 mg, 2.90 mmol) inTHF (10 mL) is added a solution of tetra-butylammonium fluoride (17.4 mLof 1 M in THF, 17.4 mmol). The reaction mixture is stirred at roomtemperature for 2 hr. The THF is removed under reduced pressure. Theresidue is dissolved in EtOAc (200 mL). The organic layer is washed withbrine then dried (Na₂SO₄), filtered and concentrated in vacuo. The crudeproduct is chromatographed on silica gel, eluting with Hex:EtOAc (3:1)to afford the title compound (480 mg, 86%).

[0162]¹H NMR (300 MHz, CD₃OD) δ 3.81 (s, 3H), 6.73 (s, 1H), 7.22 (s, 1H)

PREPARATION 26 5-Bromo-4-methoxythiophene-2-sulfonamide

[0163]

[0164] To a solution of 4-methoxythiophene-2-sulfonamide (240 mg, 1.24mmol) in CH₂Cl₂ (40 mL) is added N-bromosuccinimide (287 mg, 1.61 mmol).The reaction mixture is stirred at 0° C. for 7 hr. After 7 hr, thereaction mixture is diluted with CH₂Cl₂ (150 mL). The organic layer iswashed with brine then dried (Na₂SO₄), filtered and concentrated invacuo. The crude product is chromatographed on silica gel, eluting withHex:EtOAc (2:1) to afford the title compound (277 mg, 82%).

[0165]¹H NMR (300 MHz, CD₃OD) δ 3.30 (s, 3H), 7.40 (s, 1H)

PREPARATION 27 2-Trimethylsilyl-3-methylsulfanylthiophene

[0166]

[0167] A solution of n-BuLi (5.3 mL of 1.6 M in hexanes, 8.5 mmol) isadded dropwise to a solution of 3-methylsulfanylthiophene (1.0 g, 7.7mmol) in anhydrous Et₂O(8 mL) under nitrogen at −70° C. The mixture isstirred at −70° C. for 2 hr. Chlorotrimethylsilane (1.5 mL) is addedslowly to the reaction mixture. The mixture is warmed to roomtemperature and stirred for 3 hr. The reaction is quenched with water(50 mL) and Et₂O (50 mL). The aqueous layer is extracted with Et₂O (50mL). The combined organic extracts are dried (Na₂SO₄), filtered andconcentrated. The crude product is chromatographed on silica gel,eluting with hexanes to afford the title compound (0.75 g, 48%) as acolorless liquid.

[0168]¹H NMR 300 MHz (CD₃OD) δ 0.38 (s, 9H), 2.42 (s, 3H), 7.17 (d,J=3.7 Hz, 1H), 7.51 (d, J=3.7 Hz, 1H)

PREPARATION 28 (5-Trimethylsilyl-4-methylsulfanylthiophene-2-sulfonamide

[0169]

[0170] A solution of n-BuLi (7.4 mL of 2.5 M in hexanes, 18.4 mmol) isadded dropwise to a solution of2-trimethylsilyl-3-methylsulfanylthiophene (1.5 g, 7.4 mmol) inanhydrous THF (25 mL) under nitrogen at −70° C. The mixture is stirredat at −70° C. for 4 hr. Sulfur dioxide is bubbled through the solutionat −70° C. for 5 minutes. After 2.5 hr, N-chlorosuccinimide (1.98 g,14.8 mmol) is added to the suspension. The mixture is stirred at roomtemperature for 1 hr. The reaction mixture is filtered and solids washedwith CH₂Cl₂. The filtrate is concentrated and the residue dissolved inCH₂Cl₂ (200 mL). The organic layer is washed with brine then dried(Na₂SO₄), filtered and concentrated. The residue is dissolved in acetone(20 mL) and added to a solution of 29% NH₄OH (13 mL) in acetone (30 mL)at 0° C. The mixture is stirred at 0° C. for 30 min. The acetone isremoved under reduced pressure and the residue is extracted with EtOAc(2×100 mL). The organic extracts are washed with brine then dried(Na₂SO₄), filtered and concentrated. The crude product ischromatographed on silica gel, eluting with Hex:EtOAc (3:1) to affordthe title compound (0.65 g, 34%).

[0171]¹H NMR (300 MHz, CD₃OD) δ 0.39 (s, 9H), 2.45 (s, 3H), 7.65 (s, 1H)

PREPARATION 29 4-Methylsulfanylthiophene-2-sulfonamide

[0172]

[0173] To a solution of5-trimethylsilyl-4-methylsulfanylthiophene-2-sulfonamide (660 mg, 2.34mmol) in THF (10 mL) is added a solution of tetra-butylammonium fluoride(14.0 mL of 1 M in THF, 14.0 mmol). The reaction mixture is stirred atroom temperature for 3 hr. The THF is removed under reduced pressure andthe residue is dissolved in EtOAc (200 mL). The organic layer is washedwith brine then dried (Na₂SO₄), filtered and concentrated in vacuo. Thecrude product is chromatographed on silica gel, eluting with Hex:EtOAc(2:1) to afford the title compound (400 mg, 82%).

[0174]¹H NMR (300 MHz, CD₃OD) δ 2.49 (s, 3H), 7.35 (s, 1H), 7.47 (s, 1H)

PREPARATION 30 5-Bromo-4-methylsulfanylthiophene-2-sulfonamide

[0175]

[0176] To a solution of 4-methylsulfanylthiophene-2-sulfonamide (210 mg,1.00 mmol) in CHCl₃ (10 mL) and AcOH (10 mL) is added N-bromosuccinimide(231 mg, 1.30 mmol). The reaction mixture is stirred at room temperaturefor 7 hr. After 7 hr, the reaction mixture is neutralized with 1 M NaOHand the solution is extracted with EtOAc (200 mL). The organic layer iswashed with brine then dried (Na₂SO₄), filtered and concentrated invacuo. The crude product is chromatographed on silica gel, eluting withHex:EtOAc (3:1) to afford the title compound (200 mg, 70%).

[0177]¹H NMR (300 MHz, CD₃OD) δ 2.49 (s, 3H), 7.45 (s, 1H)

PREPARATION 31 2,4-Dibromobenzonitrile

[0178] Copper cyanide (2.32 g, 25.9 mmol) is added to stirred anhydrousdimethylsulfoxide (50 mL) at 60° C. to form a clear solution, followedby the addition of tert-butylnitrite (7.1 mL, 59.7 mmol) all at once. Asolution of 2,4-dibromoaniline 21 (5.0 g, 19.9 mmol) in anhydrousdimethylsulfoxide (30 mL) is added dropwise, via cannula, to themixture. After the addition is complete, the reaction mixture is allowedto stir for 1 hr. After being cooled to 45° C., the mixture is slowlytreated with 5N hydrochloric acid (50 mL). Five minutes later, thereaction mixture is cooled to ambient temperature before being extractedwith ethyl acetate/hexane (1:1; 2×300 mL). The combined organic layersare washed with water (100 mL) and brine (100 mL), dried, concentratedin vacuo, and then chromatographed on silica (0-5% ethyl acetate inhexane) to give the title compound (1.61 g, 31% yield). FD(+)MS m/z 259,(M⁺) consistent with 2 Br.

PREPARATION 32 2,4-Dibromobenzoic acid

[0179] A stirred suspension of 2,4-dibromobenzonitrile (1.57 g, 6.0mmol) in sulfuric acid (6 M, 150 mL) is heated to reflux for 3 days. Thereaction mixture is cooled to ambient temperature before being extractedwith ethyl acetate (2×75 mL). The combined organic layers are washedwith water (100 mL) and brine (50 mL), dried, concentrated, and thenchromatographed on silica (acetic acid/methyl alcohol/chloroform,0.1:0.5:99.4) to give the title compound (0.81 g, 48% yield). mp171-172° C.; ES(−)MS m/z 277, (M−H)⁻ consistent with 2 Br.

PREPARATION 33 2-Bromo-4-chlorobenzoic acid

[0180]

[0181] An aqueous solution of sodium nitrate (2.21 g) in water (15 mL)is added dropwise to a stirred, ice-cooled mixture of2-amino-4-chlorobenzoic acid (5.00 g, 29.1 mmol) and 48% hydrobromicacid (150 mL) in water (150 mL). The resultant mixture is stirred for 2hr at 0° C. Then it is treated dropwise with an aqueous solution ofcopper bromide (7.81 g) in water (20 mL). Upon the completion of theaddition, the reaction mixture is allowed to warm to ambient temperaturewhere it is stirred overnight. After extraction with ethylacetate/hexanes (3:1; 2×400 mL), the combined organic layers are washedwith brine (200 mL), dried, concentrated, and chromatographed on silica(1% methyl alcohol and 0.5% acetic acid in chloroform) to give the titlecompound (4.04 g, 59% yield). mp 154-155° C.; ES(−)MS m/z 233, (M−H)⁻consistent with 1 Br and 1 Cl.

PREPARATION 34 2-Chloro-4-methylbenzoic acid

[0182]

[0183] To 4-bromo-3-chlorotoluene (4.97 g, 24.2 mmol) indimethylformamide (25 mL) is added palladium acetate (0.54 g, 2.42mmol), 1,3-bis(diphenylphosphino)propane (0.998 g, 2.42 mmol),triethylamine (12.5 mL) and methanol (12.5 mL). The reaction vessel isevacuated and purged three times with carbon monoxide gas. A balloonfilled with carbon monoxide gas is used to maintain the carbon monoxideatmosphere. The reaction mixture is heated at 80° C. for 8 hr. Themixture is washed with water and extracted with hexanes (2×50 mL). Thecombined organic layers are dried over sodium sulfate, filtered,concentrated, and chromatographed with 0-3% ethyl acetate in hexanes.1.24 g (28%) of methyl 2-chloro-4-methylbenzoate is isolated as acolorless oil.

[0184] ES(+)MS m/z 184, (M+H)⁺ consistent with 1 Cl.

[0185] To methyl 2-chloro-4-methylbenzoate (1.00 g, 5.42 mmol) intetrahydrofuran (10 mL) methyl alcohol (5 mL) and water (2.5 mL) isadded 2N lithium hydroxide (8.12 mL, 16.2 mmol). The reaction mixture isheated at 50° C. for 2.5 hr, cooled to room temperature, and thenquenched with 5N hydrochloric acid (3.24 mL). The mixture isconcentrated to remove the tetrahydrofuran and methyl alcohol. A whiteprecipitate is formed and is filtered. After drying, 0.922 g (100%) of2-chloro-4-methylbenzoic acid is isolated. ES(−)MS m/z 169, (M−H)⁻consistent with 1 Cl.

PREPARATION 35 4,4,4-Trifluoro-3-methoxy-but-2-enoic acid ethyl ester

[0186] To a solution of ethyl 4,4,4-trifluoroacetoacetate (12 mL, 82mmol) in DMF (80 mL) is added cesium carbonate (26.4 g, 82 mmol). Thereaction mixture is heated to 70° C. A solution of methylp-toluenesulfonate (13.5 mL, 90 mmol) in DMF (30 mL) is then addeddropwise during 30 min and the reaction mixture is stirred for anadditional 1 hr. After cooling to room temperature, the reaction mixtureis diluted with H₂O (150 mL) and extracted with Et₂O (2×150 mL). Theorganic extracts are combined and washed with H₂O and brine, then dried(Na₂SO₄), filtered and concentrated to afford the title compound (9.0 g,56%) as an oil which is used without further purification.

[0187]¹H NMR (300 MHz, CDCl₃) δ 1.28 (t, J=7.1 Hz, 3H), 4.01 (s, 3H),4.19 (q, J=7.1 Hz, 2H), 5.75 (s, 1H)

PREPARATION 36 3-Hydroxy-5-trifluoromethyl-thiophene-2-carboxylic acidmethyl ester

[0188] A solution of 4,4,4-trifluoro-3-methoxy-but-2-enoic acid ethylester (9.6 g, 48.5 mmol) and methyl thioglycolate (4.3 mL, 48.5 mmol) inMeOH (75 mL) is cooled to 5° C. A solution of KOH (3.3 g, 58.2 mmol) inMeOH (75 mL) is then added over 30 min. The reaction mixture is stirredovernight at room temperature. The reaction mixture is then poured overa stirred mixture of ice (75 g), H₂O (75 mL) and concentrated H₂SO₄ (4.5mL). The mixture is extracted with EtOAc (2×250 mL). The combinedextracts are washed with saturated NaHCO₃. The washings areback-extracted with EtOAc. The combined organic layers are washed withbrine, then dried (Na₂SO₄), filtered and concentrated to afford thetitle compound (10 g, 91%) as a brown oil which is used without furtherpurification.

[0189]¹H NMR (300 MHz, CDCl₃) δ 3.92 (s, 3H), 7.06 (s, 1H), 9.48 (br s,1H)

PREPARATION 37 3-Hydroxy-5-trifluoromethyl-thiophene-2-carboxylic acid

[0190] To a stirred solution of NaOH (8.0 g, 200 mmol) in H₂O (25 mL) isadded a solution of 3-hydroxy-5-trifluoromethyl-thiophene-2-carboxylicacid methyl ester (11.4 g, 50 mmol) in MeOH (25 mL). The reactionmixture is heated at reflux for 3 hr and then cooled to roomtemperature. The reaction mixture is concentrated to about half volumeand cooled to 5° C. Acidification to pH 1 with concentrated HCl (17 mL)results in a suspension. After stirrng the suspension for 30 min at 5°C., the solids are collected by filtration, washed with H₂O and driedunder vacuum to afford the sub-title compound (8.5 g, 79%) as anoff-white solid which is used without further purification.

[0191]¹H NMR (300 MHz, CDCl₃) δ 7.30 (s, 1H), 11.7 (br s, 2H)

PREPARATION 38 5-Trifluoromethyl-thiophen-3-ol

[0192] 3-Hydroxy-5-trifluoromethyl-thiophene-2-carboxylic acid (8.0 g,37.8 mmol) is placed in a flask and heated to 105° C. under argon.Heating is continued for 2 hr to complete the decarboxylation. Uponcooling, the title compound is obtained (6.8 g, 85%) as a brown oilwhich is used without further purification.

[0193]¹H NMR (300 MHz, CDCl₃) enol (major) δ 5.01 (br s, 1H), 6.52 (d,J=1.7 Hz), 7.06 (m, 1H)

[0194]¹H NMR (300 MHz, CDCl₃) keto (minor) δ 3.86 (s, 2H), 6.59 (br s,1H)

PREPARATION 391-Phenyl-5-(5-trifluoromethyl-thiophen-3-yloxy)-1H-tetrazole

[0195] A solution of 5-trifluoromethyl-thiophen-3-ol (2.0 g, 11.9 mmol)in dry acetone (480 mL) containing 5-chloro-1-phenyl-1H-tetrazole (2.1g, 11.9 mmol) and K₂CO₃ (3.3 g, 23.8 mmol) is maintained at reflux withcareful exclusion of moisture overnight. The acetone is removed underreduced pressure and the residue is partitioned between CH₂Cl₂ (500 mL)and H₂O (50 mL). The organic extracts are washed with brine, then dried(Na₂SO₄), filtered and concentrated. The crude product ischromatographed on silica gel, eluting with EtOAc:Hex (1:80) to affordthe title compound (2.5 g, 68%) as a white solid.

[0196]¹H NMR (300 MHz, CDCl₃) δ 7.52-7.61 (m, 4H), 7.73 (d, J=7.7 Hz,2H), 7.79 (s, 1H)

PREPARATIONS 40 AND 413-(1-Phenyl-1H-tetrazol-5-yloxy)-5-trifluoromethyl-thiophene-2-sulfonamideand3-[1-(4-Sulfamoyl-phenyl)-1H-tetrazol-5-yloxy]-5-trifluoromethyl-thiophene-2-sulfonamide

[0197] A solution of chlorosulfonic acid (2 mL, 30 mmol) is placed in aflask and 1-phenyl-5-(5-trifluoromethyl-thiophen-3-yloxy)-1H-tetrazole(100 mg, 0.30 mmol) is added to the solution under a nitrogenatmosphere. The solution was heated to 100° C. for 2 hr. The solution iscooled to 70° C. and thionyl chloride (0.1 mL, 0.33 mmol) is added thenthe reaction is reheated to 100° C. and stirred for an additional 2 hr.The reaction mixture is poured onto ice dropwise and the solution isextracted with CH₂Cl₂ (100 mL). The organic layer is washed with brine,then dried (Na₂SO₄), filtered and concentrated. The residue is dissolvedin acetone (5 mL) and is added to a solution of 29% NH₄OH (5 mL) andacetone (10 mL) at 0° C. The mixture is stirred at 0° C. for 30 min. Theacetone is removed under reduced pressure and the residue is extractedwith EtOAc (2×50 mL). The organic extracts are washed with brine, thendried (Na₂SO₄), filtered and concentrated. The crude product ischromatographed on silica gel, eluting with EtOAc:Hex (1:3) to afford amixture of the title compounds (91 mg, 65%) as a white solid. In anotherreaction, the components are separated by chromatography on silica geleluting with EtOAc:Hex (1:5) and characterized individually.

[0198]¹H NMR (300 MHz, CD₃OD) δ 7.57-7.67 (m, 4H), 7.89 (d, J=5.9 Hz,2H)

[0199]¹H NMR (300 MHz, CD₃OD) δ 7.96 (d, J=4.2 Hz, 1H), 8.15 (s, 4H)

PREPARATION 42 5-Trifluoromethyl-thiophene-2-sulfonamide

[0200] To a solution of3-[1-(4-sulfamoyl-phenyl)-1H-tetrazol-5-yloxy]-5-trifluoromethyl-thiophene-2-sulfonamide(210 mg, 0.47 mmol) in benzene (50 mL) is added H₂O (2 mL), EtOH (3 mL),formic acid (2 mL) and 10% palladium on carbon (350 mg). The mixture isheated to 80° C. overnight. The reaction mixture is cooled to roomtemperature and diluted with benzene (50 mL). The reaction mixture isfiltered. The benzene layer is dried (Na₂SO₄), filtered andconcentrated. The crude product is chromatographed on silica gel,eluting with EtOAc:Hex (1:10) to afford the title compound (18 mg, 17%)as a white solid.

[0201] The same procedure is applied to3-(1-phenyl-1H-tetrazol-5-yloxy)-5-trifluoromethyl-thiophene-2-sulfonicacid amide to also produce the title compound.

[0202]¹H NMR (300 MHz, CD₃OD) δ 7.56 (d, J=4.0 Hz, 1H), 7.60 (d, J=4.0Hz, 1H) ES(−)MS m/z 230, (M−H)⁻.

General Coupling Procedure

[0203] To a stirring solution of the benzoic acid (1.25 eq) in drydichloromethane (10 mL/mmol), the sulfonamide (1.0 eq) is added in oneportion followed by EDC (1.25-1.5 eq) and finally,N,N-[dimethyl]4-aminopyridine (1.2 eq). The mixture is vigorouslystirred under nitrogen for 16 hr, concentrated under reduced pressure,and the residue partitioned between ethyl acetate and water. The organiclayer is washed with 1N hydrochloric acid (4 times, 20 mL/mmol), thenthe combined aqueous phases extracted with ethyl acetate (twice, 20mL/mmol). The combined organic layers are finally washed with water andsaturated aqueous sodium chloride, dried over sodium sulfate, andconcentrated under reduced pressure. The residue may be subjected tosilica gel chromatography, reversed phase chromatography orcrystallization if necessary or desired.

[0204] The compounds of EXAMPLES 1-53 are prepared essentially asdescribed in the general coupling procedure. Mass Spectral Data Example# Product (m/z) 1 N-[4-bromo-2-chlorobenzoyl]- ES(−)MS m/z 412, (M − H)⁻consistent with 1 Br 5-chlorothiophene-2- and 2 Cl. sulfonamide 2N-[4-chloro-2-methylbenzoyl]- ES(−)MS m/z 392, (M − H)⁻ consistent with1 Br 5-bromothiophene-2- and 1 Cl. sulfonamide 3N-[4-bromo-2-chlorobenzoyl]- ES(−)MS m/z 490, (M − H)⁻ consistent with4-bromo-5-chlorothiophene-2- 2 Br and 2 Cl. sulfonamide 4N-[2,4-bis(trifluoromethyl)- ES(−)MS m/z 436, (M − H)⁻ consistent withbenzoyl]-5-chlorothiophene-2- 1 Cl. sulfonamide 5N-[2,4-bis(trifluoromethyl)- ES(−)MS m/z 480, (M − H)⁻ consistent withbenzoyl]-5-bromothiophene-2- 1 Br. sulfonamide 6N-[2,4-dimethylbenzoyl]-5- ES(−)MS m/z 328 (M − H)⁻ consistent withchlorothiophene-2-sulfonamide 1 Cl. 7 N-[2-chloro-4-methylbenzoyl]-ES(+)MS m/z 394 (M + H)⁺ consistent with 5-bromothiophene-2- 1 Br and 1Cl. sulfonamide 8 N-[2-chloro-4-methylbenzoyl]- ES(+)MS m/z 350, (M +H)⁺ consistent with 5-chlorothiophene-2- 2 Cl sulfonamide 9N-[4-chloro-2-fluorobenzoyl]- ES(−)MS m/z 396, (M − H)⁻ consistent with5-bromothiophene-2- 1 Br and 1 Cl sulfonamide 10N-[2-bromo-4-methylbenzoyl]-5- ES(−)MS m/z 438, (M + H)⁺ consistent withbromothiophene-2-sulfonamide 2 Br. 11 N-[2-bromo-4-methylbenzoyl]-5-ES(+)MS m/z 394, (M + H)⁺ consistent with chlorothiophene-2-sulfonamide1 Br and 1 Cl. 12 N-[4-methyl-2-trifluoromethyl- ES(−)MS m/z 382, (M −H)⁻ consistent with benzoyl]-5-chlorothiophene-2- 1 Cl. sulfonamide 13N-[2,4-dichlorobenzoyl]-5- ES(−)MS m/z 380, (M − H)⁻ consistent with(methylthio)thiophene-2- 2 Cl. sulfonamide 14N-[4-chloro-2-methylbenzoyl]-5- ES(−)MS m/z 360, (M − H)⁻ consistentwith (methylthio)thiophene-2- 1 Cl. sulfonamide 15N-[4-methyl-2-bromobenzoyl]-5- ES(−)MS m/z 404, (M − H)⁻ consistent with(methylthio)thiophene-2- 1 Br. sulfonamide 16 N-[2,4-dichlorobenzoyl]-5-ES(−)MS m/z 348, (M − H)⁻ consistent with (methyl)thiophene-2- 2 Cl.sulfonamide 17 N-[2,4-dichlorobenzoyl]-5- ES(−)MS m/z 362, (M − H)⁻consistent with (ethyl)thiophene-2-sulfonamide 2 Cl. 18N-[2,4-dichlorobenzoyl]-5- ES(−)MS m/z 376, (M − H)⁻ consistent with(propyl)thiophene-2-sulfonamide 2 Cl. 19 N-[2,4-dichlorobenzoyl]-5-ES(−)MS m/z 364, (M − H)⁻ consistent with methoxythiophene-2- 2 Cl.sulfonamide 20 N-[2,4-dichlorobenzoyl]-5- ES(−)MS m/z 378 (M − H)⁻consistent with 2 Cl. methoxymethyl-thiophene-2- sulfonamide 21N-[2-methyl-4-bromobenzoyl]-4- ES(−)MS m/z 436, (M − H)⁻ consistent withbromothiophene-2-sulfonamide 2 Br. 22 N-[2-methyl-4-chlorobenzoyl]-2-ES(−)MS m/z 349, (M − H)⁻ consistent with chlorothiazole-5-sulfonamide 2Cl. 23 N-[2,4-dichlorobenzoyl]-2- ES(−)MS m/z 369, (M − H)⁻ consistentwith chlorothiazole-5-sulfonamide 3 Cl. 24 N-[2,4-dichlorobenzoyl]-2-ES(−)MS m/z 365, (M − H)⁻ consistent with methoxythiazole-5-sulfonamide2 Cl. 25 N-[2-methyl-4-chlorobenzoyl]-2- ES(−)MS m/z 345, (M − H)⁻consistent with methoxythiazole-5-sulfonamide 1 Cl. 26N-[2,4-dichlorobenzoyl]-4,5- ES(−)MS m/z 490, (M − H)⁻ consistent withdibromothiophene-2- 1 Br and 2 Cl. sulfonamide 27N-[4-bromo-2-methylbenzoyl]- ES(−)MS m/z 514, (M − H)⁻ consistent with4,5-dibromothiophene-2- 3 Br. sulfonamide 28N-[4-chloro-2-methylbenzoyl]-5- ES(−)MS m/z 341, (M + H)⁺ consistentwith cyanothiophene-2-sulfonamide 1 Cl. 29N-[4-bromo-2-methylbenzoyl]-5- ES(+)MS m/z 385, (M + H)⁺ consistent withcyanothiophene-2-sulfonamide 1 Br. 30 N-[4-chloro-2-methylbenzoyl]-5-ES(+)MS m/z 350, (M + H)⁺ consistent with chlorothiophene-2-sulfonamide2 Cl. 31 N-[2-bromo-4-methylbenzoyl]-5- ES(−)MS m/z 392, (M − H)⁻consistent with chlorothiophene-2-sulfonamide 1 Br and 1 Cl. 32N-[2,4-dibromobenzoyl]-5- ES(−)MS m/z 500, (M − H)⁻ consistent withbromothiophene-2-sulfonamide 3 Br. 33 N-[2-bromo-4-chlorobenzoyl]-5-ES(−)MS m/z 456, (M − H)⁻ consistent with 2 bromothiophene-2-sulfonamideBr and 1 Cl. 34 N-[2-methyl-4-bromobenzoyl]-4- ES(−)MS m/z 392, (M − H)⁻consistent with chlorothiophene-2-sulfonamide 1 Br and 1 Cl. 35N-[2,4-dichlorobenzoyl]-4- ES(−)MS m/z 368, (M − H)⁻ consistent withchlorothiophene-2-sulfonamide 3 Cl. 36 N-[2,4-dichlorobenzoyl]-4-ES(−)MS m/z 446, (M − H)⁻ consistent with chloro-5-bromothiophene-2- 1Br and 3 Cl. sulfonamide 37 N-[2,4-dichlorobenzoyl]-4- ES(−)MS m/z 426,(M − H)⁻ consistent with methyl-5-bromothiophene-2- 1 Br and 2 Cl.sulfonamide 38 N-[2,4-dichlorobenzoyl]-4- ES(−)MS m/z 348, (M − H)⁻consistent with methylthiophene-2-sulfonamide 2 Cl. 39N-[2-methyl-4-bromobenzoyl]-4- ES(−)MS m/z 388, (M − H)⁻ consistent withmethoxythiophene-2- 1 Br. sulfonamide 40 N-[2,4- ES(−)MS m/z 416, (M −H)⁻. bistrifluoromethylbenzoyl]-4- methylthiophene-2-sulfonamide 41N-[2,4-dichlorobenzoyl]-4- ES(−)MS m/z 364, (M − H)⁻ consistent withmethoxythiophene-2- 2 Cl. sulfonamide 42 N-[2-methyl-4-bromobenzoyl]-4-ES(−)MS m/z 404, (M − H)⁻ consistent with methylthio-thiophene-2- 1 Br.sulfonamide 43 N-[2,4-dichlorobenzoyl]-4- ES(−)MS m/z 380, (M − H)⁻consistent with methylthio-thiophene-2- 2 Cl. sulfonamide 44 N-[2,4-ES(−)MS m/z 432, (M − H)⁻. bistrifluoromethylbenzoyl]-4-methoxythiophene-2- sulfonamide 45 N-[2,4- ES(−)MS m/z 448, (M − H)⁻.bis(trifluoromethyl)benzoyl]-4- methylthio-thiophene-2- sulfonamide 46N-[2,4-dichlorobenzoyl]-4- ES(−)MS m/z 458, (M − H)⁻ consistent withmethylthio-5-bromothiophene-2- 1 Br and 2 Cl sulfonamide 47N-[2,4-dichlorobenzoyl]-4- ES(−)MS m/z 442, (M − H)⁻ consistent withmethoxy-5-bromothiophene-2- 1 Br and 2 Cl sulfonamide 48N-[2-methyl-4-bromobenzoyl]-4- ES(−)MS m/z 466, (M − H)⁻ consistent withmethoxy-5-bromothiophene-2- 2 Br. sulfonamide 49N-[2-methyl-4-bromobenzoyl]-4- ES(−)MS m/z 482, (M − H)⁻ consistent withmethylthio-5-bromothiophene-2- 2 Br. sulfonamide 50N-[2,4-dichlorobenzoyl]-2- ES(−)MS m/z 377, (M − H)⁻ consistent withisopropylthiazole-5-sulfonamide 2 Cl. 51 N-[2-methyl-4-bromobenzoyl]-2-ES(−)MS m/z 401, (M − H)⁻ consistent withisopropylthiazole-5-sulfonamide 1 Br. 52 N-[2-methyl-4-bromobenzoyl]-2-ES(−)MS m/z 373, (M− H)⁻ consistent with methylthiazole-5-sulfonamide 1Br. 53 N-[2,4-dichloro-benzoyl]-5- ES(−)MS m/z 402, (M − H)⁻ consistentwith 2 trifluoromethylthiophene-2- Cl. sulfonamide

EXAMPLE 54 N-[4-bromo-2-chlorobenzoyl]-5-bromothiophene-2-sulfonamide

[0205]

[0206] An 8 mL reaction vial is charged with 4-bromo-2-chlorobenzoicacid (0.39 mmol, 1.5 eq) and 2.0 mL of dichloromethane. A stock solution(4.0 mL) containing 5-bromothiophene-2-sulfonamide (0.26 mmol, 1 eq) andN,N-[dimethyl]-4-aminopyridine (48 mg, 0.39 mmol, 1.5 eq) indichloromethane is added, followed by 0.261 g carbodiimide polystyreneresin (2.0 mmol/g, 0.52 mmol, 2.0 eq, Novabiochem) and the vial iscapped and shaken. After 72 hr, 0.77 g sulphonated polystyrene resin(MP-TsOH) is added (1.53 mmol/g, 1.17 mmol, Argonaut). After about 18 hrthe reaction mixture is filtered and concentrated under reducedpressure. Chromatography was applied to the residue and fractionscontaining product were combined and concentrated under reduce pressureto provide the title compound.

[0207] ES(−)MS m/z 456, (M−H)⁻ consistent with 2 Br and 1 Cl.

[0208] The compounds of Examples 55-62 are prepared essentially asdescribed in Example 54. Mass Spectral Data Example # Product (m/z) 55N-[2,4-dichlorobenzoyl]-thiophene-2- ES(−)MS m/z 334, (M − H)⁻sulfonamide consistent with 2 Cl. 56N-[2,4-dichlorobenzoyl]-5-(2-pyridyl)- ES(−)MS m/z 411, (M − H)⁻thiophene-2-sulfonamide consistent with 2 Cl. 57N-[4-bromo-2-methylbenzoyl]-5- ES(−)MS m/z 436, (M − H)⁻bromothiophene-2-sulfonamide consistent with 2 Br. 58N-[2-chloro-4-nitrobenzoyl]-5-bromothiophene- ES(−)MS m/z 423, (M − H)⁻2-sulfonamide consistent with 1 Br and 1 Cl. 59N-[2,4-dimethylbenzoyl]-5-bromothiophene-2- ES(−)MS m/z 372, (M − H)⁻sulfonamide consistent with 1 Br. 60 N-[4-chloro-2-methylbenzoyl]-5-ES(−)MS m/z 348, (M − H)⁻ chlorothiophene-2-sulfonamide consistent with2 Cl. 61 N-[2,4-dichlorobenzoyl]-5-chlorothiophene-2- ES(−)MS m/z 368,(M − H)⁻ sulfonamide consistent with 3 Cl. 62 N-[2,4-dichlorobenzoyl]-5-ES(−)MS m/z 442, (M − H)⁻ (phenylthio)thiophene-2-sulfonamide consistentwith 2 Cl.

EXAMPLE 63 N-[2,4-dichlorobenzoyl]-5-bromothiophene-2-sulfonamide

[0209]

[0210] To a reaction mixture of dichlorobenzoic acid (28.4 g, 148.7mmol), 5-bromo-2-sulfonamide (30.0 g, 123.9 mmol) and EtOAc (200.0 mL)at room temperature is added a hot solution of CDI (24.1 g, 148.7 mmol)in THF (100.0 mL) over a period of 13.0 min. Extra THF (50.0 mL) isadded to aid and wash residual CDI into the reaction vessel. Gasevolution is observed during addition of CDI solution/slurry. This canbe controlled by the rate of addition. At the end of CDI addition, thelight yellow solution is stirred for 10 min, and then heated at refluxfor 90 min or until no gas evolution is observed (reaction intermediateis monitored by GC and deemed complete when no acid peak is observed).The reaction is then allowed to equilibrate to 40° C. after which neatDBU (22.3 mL, 148.7 mm) is added all at once (maximum temperatureattained by the end of addition is 45° C.) and stirred to roomtemperature overnight for convenience. The reaction is deemed completeby HPLC with the disappearance of the sulfonamide starting material.Deionized water (250.0 mL) is then added and the top organic layerseparated. The aqueous layer is back extracted with EtOAc (50. mL). Thecombined organic layers are washed vigorously with 1N HCl solution(500.0 ml), dried with anhydrous MgSO₄, filtered and the cake washedwith EtOAc (20.0 mL). The filtrate is then concentrated at reducedpressure (water bath temp. 50° C.) to 70.4 g of a thick solution. Tothis solution is added heptane (200.0 mL) with vigorous stirring untilan off-white precipitate forms in about an hour. The precipitate isfiltered and the cake washed with heptane (25.0 mL). The precipitate isthen dried in a house vacuum at 55° C. for 18 hr (45.4 g, 88.2% wtyield). ES(−)MS m/z 412, (M−H)⁻ consistent with 1 Br and 2 Cl.

EXAMPLE 64 N-[2,4-dichlorobenzoyl]-5-bromothiophene-2-sulfonamide sodiumsalt

[0211]

[0212] To a solution of the compound of Example 63 (25.0 g, 60.2 mmol)and MTBE (208.0 mL) at room temperature is added sodium methoxide (3.3g, 60.2 mmol) in a single portion. The reaction is then stirred for 24hr, after which heptane (426.0 ml) is added followed by vigorousstirring for 60 min. A white precipitate forms and is then filteredunder a positive nitrogen pressure, and the cake subsequently washedwith heptane (150.0 mL). The precipitate is then pulled to semi-dryness,followed by drying in a house vacuum oven at 100° C. for 18 hr(mass=22.1 g, 84% wt. Yield; ¹H NMR (DMSO d₆) 7.13-7.14 δ (d, J=3.9 Hz,1H), 7.30-7.35 (m, 2H), 7.47-7.52 (m, 2H)).

[0213] All of the compounds concerned are orally available and arenormally administered orally, and so oral administration is preferred.However, oral administration is not the only route or even the onlypreferred route. For example, transdermal administration is verydesirable for patients who are forgetful or petulant about taking oralmedicine, and the intravenous route may be preferred as a matter ofconvenience or to avoid potential complications related to oraladministration. Compounds of Formula I may also be administered by thepercutaneous, intramuscular, intranasal or intrarectal route inparticular circumstances. The route of administration may be varied inany way, limited by the physical properties of the drugs, theconvenience of the patient and the caregiver, and other relevantcircumstances (Remington's Pharmaceutical Sciences, 18th Edition, MackPublishing Co. (1990)).

[0214] The pharmaceutical compositions are prepared in a manner wellknown in the pharmaceutical art. The carrier or excipient may be asolid, semi-solid, or liquid material that can serve as a vehicle ormedium for the active ingredient. Suitable carriers or excipients arewell known in the art. The pharmaceutical composition may be adapted fororal, inhalation, parenteral, or topical use and may be administered tothe patient in the form of tablets, capsules, aerosols, inhalants,suppositories, solutions, suspensions, or the like.

[0215] The compounds of the present invention may be administeredorally, for example, with an inert diluent or capsules or compressedinto tablets. For the purpose of oral therapeutic administration, thecompounds may be incorporated with excipients and used in the form oftablets, troches, capsules, elixirs, suspensions, syrups, wafers,chewing gums and the like. These preparations preferably contain atleast 4% of the compound of the present invention, the activeingredient, but may be varied depending upon the particular form and mayconveniently be between 4% to about 70% of the weight of the unit. Theamount of the compound present in compositions is such that a suitabledosage will be obtained. Preferred compositions and preparations of thepresent invention may be determined by methods well known to the skilledartisan.

[0216] The tablets, pills, capsules, troches, and the like may alsocontain one or more of the following adjuvants: binders such aspovidone, hydroxypropyl cellulose, microcrystalline cellulose, orgelatin; excipients or diluents such as: starch, lactose,microcrystalline cellulose or dicalcium phosphate, disintegrating agentssuch as: croscarmellose, crospovidone, sodium starch glycolate, cornstarch and the like; lubricants such as: magnesium stearate, stericacid, talc or hydrogenated vegetable oil; glidants such as colloidalsilicon dioxide; wetting agents such as: sodium lauryl sulfate andpolysorbate 80; and sweetening agents such as: sucrose, aspartame orsaccharin may be added or a flavoring agent such as: peppermint, methylsalicylate or orange flavoring. When the dosage unit form is a capsule,it may contain, in addition to materials of the above type, a liquidcarrier such as polyethylene glycol or a fatty oil. Other dosage unitforms may contain other various materials that modify the physical formof the dosage unit, for example, as coatings. Thus, tablets or pills maybe coated with sugar, hydroxypropyl methylcellulose, polymethacrylates,or other coating agents. Syrups may contain, in addition to the presentcompounds, sucrose as a sweetening agent and certain preservatives, dyesand colorings and flavors. Materials used in preparing these variouscompositions should be pharmaceutically pure and non-toxic in theamounts used.

[0217] Injections for parenteral administration include sterile aqueousor non-aqueous solutions, suspensions and emulsions. Aqueous solutionsand suspensions may include distilled water for injection orphysiological salt solution. Non-aqueous solutions and suspensions mayinclude propylene glycol, polyethylene glycol, vegetable oil such asolive oil, alcohol such as ethanol or POLYSORBATE80 (registered trademark). Injections may comprise additional ingredients other than inertdiluents: e.g. preserving agents, wetting agents, emulsifying agents,dispersing agents, stabilizing agents (such as lactose), assistingagents such as agents to assist dissolution (e.g. glutamic acid oraspartic acid). They may be sterilized for example, by filtrationthrough a bacteria-retaining filter, by incorporation of sterilizingagents in the compositions or by irradiation. They may also bemanufactured in the form of sterile solid compositions that may bedissolved in sterile water or some other sterile diluent(s) forinjection immediately before use.

[0218] The compounds of Formula I are generally effective over a widedosage range. For example, dosages per day normally fall within therange of about 10 to about 300 mg/kg of body weight. In some instancesdosage levels below the lower limit of the aforesaid range may be morethan adequate, while in other cases still larger doses may be employedwithout causing any harmful side effect, and therefore the above dosagerange is not intended to limit the scope of the invention in any way. Itwill be understood that the amount of the compound actually administeredwill be determined by a physician, in the light of the relevantcircumstances, including the condition to be treated, the chosen routeof administration, the actual compound or compounds administered, theage, weight, and response of the individual patient, and the severity ofthe patient's symptoms.

[0219] Inhibition of HUVEC Proliferation

[0220] Human umbilical vein endothelial cells (HUVEC;BioWhittaker/Clonetics, Walkersville, Md.) were maintained inendothelial cell growth medium (EGM) containing basal medium (EBM) withbovine brain extract, human epidermal growth factor, hydrocortisone,gentamicin, amphotericin B and 2% fetal bovine serum. For the assay,HUVEC (5×10³) in EBM (200 W) with 0.5% fetal bovine serum were added towells in a 96-well cell culture plate and incubated at 37° C. for 24 hrin humidified 5% carbon dioxide/air. The test compounds were seriallydiluted in dimethyl sulfoxide (DMSO) in concentrations from 0.0013 to 40μM and added to the wells in 20 pd. Then human vascular endothelialgrowth factor (VEGF) (20 ng/ml in wells; R&D Systems, Minneapolis,Minn.) prepared from a stock solution of 100 μg/ml in phosphate bufferednormal saline containing 0.1% bovine serum albumin, was added to thewells. The HUVEC were incubated at 37° C. for 72 hr in humidified 5%carbon dioxide/air. WST-1 cell proliferation reagent (20 μl; BoehringerMannheim, Indianapolis, Ind.) was added to the wells and the platesreturned to the incubator for 1 hr. The absorbance of each well at 440nm was measured. The growth fraction was determined from the absorbanceof treated wells with and without VEGF divided by the absorbanceobtained from control wells set to zero and 1.0. The exemplifiedcompounds were tested in this assay and all exhibited an IC₅₀≦1.0 μM.

[0221] HCT116 Colon Carcinoma Cell Growth Inhibition

[0222] Human HCT116 colon carcinoma cells were grown monolayer culturein RPMI 1640 medium supplemented with 10% fetal bovine serum and 1%penicillin-streptomycin (GibcoBRL, Grand Island, N.Y.). HCT116 cells inexponential growth phase were exposed to various concentrations of thetest compounds at 37° C. for 72 hr in 5% carbon dioxide/air. Afterexposure to the agent, the cells were washed with 0.9% phosphatebuffered saline. Growth inhibition was determined using WST-1 cellproliferation reagent as described above. The results are expressed asthe growth fraction of treated cells compared with control cultures.Representative compounds of the present invention were tested forefficacy against the human colon HCT116 tumor cells. The data from theseexperiments are summarized in TABLE I. TABLE I Human Colon HCT116 tumorcells EXAMPLE IC₅₀ (μM) 1 5.6 2 6.0 3 14.7 4 7.7 6 20.6 7 5.2 9 21.7 163.7 17 5.0 18 13.2 19 5.8 20 5.7 28 8.0 29 17.3 30 15.8 31 9.1 32 3.9 5417.0 55 4.5 56 5.4 57 3.4 58 5.2 61 1.0 63 1.3

[0223] Conventional Murine Tumor and Human Tumor Xenograft Assays

[0224] Inhibition of tumors transplanted into mice is an acceptedprocedure for studying the efficacy of antitumor agents (Corbett et al.,In vivo Methods for Screening and Preclinical Testing; Use of rodentsolid tumors for drug discovery. In: Anticancer Drug Development Guide:Preclinical Screening, Clinical Trials, and Approval, B. Teicher (ed),Humana Press Inc., Totowa, N.J., Chapter 5, pages 75-99 (1997);(Corbett, et al., Int. J. Pharmacog., 33, Supplement, 102-122 (1995)).Murine tumors or human xenografts were implanted essentially asdescribed by Corbett in In vivo Methods for Screening and PreclinicalTesting: Use of rodent solid tumors for drug discovery. Briefly, themurine tumor or human xenograft was implanted subcutaneously usingeither 12-gauge trocar implants or counted number of cells. The locationfor the trocar insertion is midway between the axillary and inguinalregion along the side of the mouse. The trocar is slipped approximately¾ of an inch subcutaneously up toward the axilla before discharging thetumor fragment, and pinching the skin as the trocar is removed.Alternatively, human tumor cells prepared from cell culture (1×10⁷cells) mixed with an equal volume of Matrigel (Becton-Dickinson) wereimplanted subcutaneously in a hind-leg of a male or female nude mouse(Charles River). A test compound in vehicle or vehicle alone wasadministered by intravenous bolus injection (iv), intraperitonealinjection (ip), or oral gavage (po). Each treatment group, as well as agroup of untreated control animals, consisted of eight to ten animalsper group in each experiment. Subcutaneous tumor response was monitoredby tumor volume measurement performed twice each week over the course ofthe experiment (60-120 days). Body weights were taken as a generalmeasure of toxicity. The subcutaneous tumor data were analyzed bydetermining the median tumor weight for each treatment group over thecourse of the experiment and calculating the tumor growth delay as thedifference in days for the treatment versus the control tumors to reacha volume of either 500 or 1000 mm³.

[0225] The compound of Example 64 was tested in two separatelaboratories against a variety of murine and human tumors substantiallyas described supra. The data from these tests are summarized in TABLEII. The parameters measured in each experiment are summarized in thefollowing paragraphs.

[0226] Tumor Weight(mg)=(a×b²)/2 where a=tumor length (mm) and b=tumorwidth (mm).

[0227] Tumor Growth Delay=T−C where T is the median time (days) requiredfor the treatment group tumors to reach a predetermined size, and C isthe median time (days) for the control group tumors to reach the samesize. TABLE II Human Colon Carcinoma HT-29 Example 64 Dose (mg/kg) TumorGrowth Delay (d) Experiment A 30 0 +/− 2 60 2 +/− 2 80 2 +/− 2Experiment B 30 9 +/− 4 60 3 +/− 4 80   8 +/− 3.6

[0228] After palpable tumors were observed drug was administered IV for5 consecutive days, animals rested for 2 days and compound dosed IVagain for 5 consecutive days.

1. A compound of formula I:

where: —X═Y— is

R¹ is selected from the group consisting of halo, C₁-C₆ alkyl, and CF₃;R² is selected from the group consisting of halo, —NO₂, C₁-C₆ alkyl, andCF₃; R³ is H, C₁-C₆ alkyl, C₁-C₄ alkoxy, C₁-C₆ alkylthio, or halo; R⁴ isselected from the group consisting of H, halo, C₁-C₄ alkoxy, C₁-C₆alkyl, —COO(C₁-C₆ alkyl), C₁-C₆ alkyl optionally substituted with C₁-C₄alkoxy, cyano, C₁-C₆ alkylthio, CF₃, S-phenyl, and pyridinyl; R⁵ ishalo, C₁-C₆ alkyl, or C₁-C₄ alkoxy; or a pharmaceutically acceptablebase addition salt thereof.
 2. The compound of claim 1 wherein R¹ and R²are independently halo or C₁-C₆ alkyl.
 3. The compound of claim 1wherein R¹ and R² are both chloro or bromo, or R¹ is methyl and R² ischloro.
 4. The compound of claim 1 wherein —X═Y— is


5. The compound of claim 4 wherein R³ is selected from H, chloro, bromo,methyl, methoxy and methylthio.
 6. The compound of claims 4 wherein R⁴is selected from H, chloro, bromo, methyl, ethyl, propyl, methylthio,CH₂OCH₃, methoxy, cyano, S-phenyl and pyridinyl.
 7. The compound ofclaim 1 which is N-[2,4-dichlorobenzoyl]-5-bromothiophene-2-sulfonamideor a pharmaceutically acceptable base addition salt thereof.
 8. Thecompound of claim 1 which isN-[4-chloro-2-methyl-benzoyl]-5-chlorothiophene-2-sulfonamide or a baseaddition salt thereof.
 9. The compound of any of claim 1 wherein thepharmaceutically acceptable base addition salt is a sodium salt.
 10. Thecompound of claim 1 which isN-[2,4-dichlorobenzoyl]-5-bromothiophene-2-sulfonamide sodium salt. 11.A method of treating susceptible neoplasms in a mammal comprisingadministering to a mammal in need of such treatment an oncolyticallyeffective amount of a compound of Formula I:

where: —X═Y— is

R¹ is selected from the group consisting of halo, C₁-C₆ alkyl, and CF₃;R is selected from the group consisting of halo, —NO₂, C₁-C₆ alkyl, andCF₃; R³ is H, C₁-C₆ alkyl, C₁-C₄ alkoxy, C₁-C₆ alkylthio, or halo; R⁴ isselected from the group consisting of H, halo, C₁-C₄ alkoxy, C₁-C₆alkyl, —COO(C₁-C₆ alkyl), C₁-C₆ alkyl optionally substituted with C₁-C₄alkoxy, cyano, C₁-C₆ alkylthio, CF₃, S-phenyl, and pyridinyl; R⁵ ishalo, C₁-C₆ alkyl, or C₁-C₄ alkoxy; or a pharmaceutically acceptablebase addition salt thereof.
 12. A pharmaceutical formulation comprisinga compound of Formula I:

where: —X═Y— is

R¹ is selected from the group consisting of halo, C₁-C₆ alkyl, and CF₃;R² is selected from the group consisting of halo, —NO₂, C₁-C₆ alkyl, andCF₃; R³ is H, C₁-C₆ alkyl, C₁-C₄ alkoxy, C₁-C₆ alkylthio, or halo; R⁴ isselected from the group consisting of H, halo, C₁-C₄ alkoxy, C₁-C₆alkyl, —COO(C₁-C₆ alkyl), C₁-C₆ alkyl optionally substituted with C₁-C₄alkoxy, cyano, C₁-C₆ alkylthio, CF₃, S-phenyl, and pyridinyl; R⁵ ishalo, C₁-C₆ alkyl, or C₁-C₄ alkoxy; or a pharmaceutically acceptablebase addition salt thereof, in admixture with a pharmaceuticallyacceptable carrier or excipient.
 13. The pharmaceutical formulation ofclaim 12 comprisingN-[2,4-dichlorobenzoyl]-5-bromothiophene-2-sulfonamide or apharmaceutically acceptable base addition salt.
 14. The pharmaceuticalformulation of claim 13 comprisingN-[2,4-dichlorobenzoyl]-5-bromothiophene-2-sulfonamide sodium salt. 15.(cancelled)
 16. (cancelled)
 17. The method of claim 11 in which thesusceptible neoplasm is a tumor of the colon or rectum.